((9-Oxoxanthene-2,6-diyl)dioxy)diacetonitriles and diacetamidoxime

ABSTRACT

The disclosure is directed to ((9-oxoxanthene-2,6diyl)dioxy)diacetonitrile and diacetamidoxime which have the formula   WHERE R1 and R2 are the same and are acetonitrile or acetamidoxime. The compounds have trichomonacidal activity in vitro.

"United States Patent [1 1 s [1 1 3,917,650

.[22] Filed: May 8, 1974 Santilli et al. v I Nov. 4, 1975 [(9-0XOXANTHENE-2,6-DIYL)DIOXY]1 diyl)dioxy]diacetonitrile and diaceta midoxime which DIACETONITRILES AND have the formula DIACETANHDOXIME [75] Inventors: Arthur A. Santilli, l-lavertown;

Anthony CL Scotese, King of Prussia, both of Pa.

[73] Assignee: American Home Products Corporation, New York, NY.

[21] App]. No.: 468,055 I 52 us. Cl. 260/335; 424/283 51 lm. cl. (:07!) 311/88 where 31 9 2 the S and are acetommle or [58] Field of Search 260/335 acetamldoxlmeg H The compounds have trichomonacidal activity in Primary Examiner-N0rma S. Milestone vitro.

Attorney, Agent, or Firm-Joseph Martin Weigman [57] ABSTRACT 2 Claims, 1 Drawing Figure The disclosure is directed to {(9-oxoxanthene-2,6-

pounds of the invention m ay be schernatically r'epre sented bywhere R and R are the same an'd are "acetonitrile or acetamidoxime. A 2; I

The specific embodiments of the: invention are {(9- oxoxanthene-2,6-diyl)dioxy]diacetonitrile and i 22'- [(9-oxoxanthene-2,6-diyl)-dioxy]diacetamidoxime. i

The preparation of these compounds may berepresented schematically by the flow diagram of FIG. 1 where X is hydrogen.

The closest known 2,732,373.

The starting material 2,6-dihydroxy 9-oxoxanthene, formula II of FIG. 1, is described in O. P. Mittal and T. R Seshodri, J. Sci. Incl. Research (India) 14B, 76 (1966) and Chemical Abstracts 49, 8932 (1955).

The compound having the structure of formula III of FIG. 1 may be prepared as follows. To a suspension of 2,6-dihydroxy-9-oxoxanthene in ethanol is added a solution of sodium in ethanol. The solution is dried, for instance by evaporation, and the residue dissolved in an inert organic solvent, such as N,N-dimethylformamide. A haloacetonitrile, preferably bromoacetonitrile, is

-?'prior en ie'us. Pat. No.

added and the mixture refluxed for Ho 31hours, preferably one hour and then dilu' ted with water. The product precipitates'and is recovered and purified, for instance by filtration and recrystallization. I

The compound having the structure of formula IV of FIG. 1 may be prepared as follows. To a mixture of a compound having the structure of formula III of FIG. 1 and hydroxylamine hydrochloride in an inert organic solvent, such as N,N-dimethylformamide, is added a base, such as sodium carbonate. The mixture is heated at a temperature of 80 to 100C for 1 to 3 hours, preferably for 2 hours on a steam bath, filtered and diluted with water. The product precipitates and is collected and purified, for instance, by filtration and recrystallization.

The compound was found to have trichomonacidal activity in vitro when evaluated as follows.

A 12.5 milligram portion of test material (as based on the active moiety) is added to 2.5 milliliter (ml) of 1 percent phosphate buffer at pH 6. Further two-fold dilutions are made in the same buffer. A l milliliter volume of each dilution is transferred, to small sterile screw cap assay tubes containing 3.8 milliliters of Diamond medium (formula furnished by American Type Culture Collection, Rockville, Md.) and 0.1 ml of calf serum. Each assay tube is inoculated with 0.1 milliliter of a 48-72 hour culture of Trichomonas vaginalis trol test material;

ing no test material is-sirnil arly inoculated. The assay tubesflzare' gently shakenand the'nl incubatedtifor .48

7 hours; at 30C. Following incubation the tube s (are, gently shaken and with the aid of a Pasteur'pipettea.

drop is deposited on a Spiers-Levy eosinophil counting chamber. The chamber is examined by phase contrast microscopy. The number of organisms present in one square millimeter isxrnultiplied by 5000 in order to express the count per milliliter. The difference between the number of organisms present, in the control tube and in the tubes containing the testr'nate rial represents the relative potency of the test material and is expressed as the percentage kill at the specific dose level. Metronidazole, may be I The compound 2-2 '[(9oxoxanthene-2 ,6 diyl)dio x y ]dia cet arni doxime showed a 33 percentkill a gram per milliliter. V I

.Whenthe compounds of this invention are employed as described above, they may be. administeredaalone or in'combination with pharmacologicallyi acceptable carriers, the proportion of which is determined by the sol-.

ubility and chemical nature .of the compound, chosen route of administration and, standard pharmacological practice. For example, they may be administered'o rally in the form of tablets or capsules containing suchexcipients as starch, milk, sugar, certain types of clay and so forth. They may be administered sublingually in the form of 'troches orlozenges in whichethei active ingredient is mixed with sugar and cornsyrups; and-then dehy-:

The dosage of 'the prese nt therapeutic varyzwith the for-in of-ad'ministrationiiind the compound chosen. Furthermore, it will vary ;partic'ul'ar subject under treatment". Generally, heatment is initiated with small dosages substantially less than the optimum dose of the compound. Thereafter the dosage is increased by small increments until the optimum effect under the circumstances is reached. It will generally be found that when the composition is administered orally, larger quantities of the active agent will be required to produce the same effect as a smaller quantity given parenterally. In general, the compounds of this invention are most desirably administered at a concentration level that will generally afford effective results without causing any harmful or deleterious side effects.

In order more clearly to disclose the nature of the present invention, specific examples of the practice of the invention are hereinafter given. It should be understood, however, that this is done solely by way of example and is intended neither to delineate the scope of the invention nor limit the ambit of the appended claims. In the examples, all temperatures are stated in degrees centigrade, and the following abbreviations are used: g" for grams, ml for milliliters, min" for minutes, and hr for hours. i

used as a suitable positive con- 1 m me,

EXAMPLE 1 The following illustrates the preparation of [(9-oxoxanthene-2,6-diyl)dioxyldiacetonitrile a compound of the formula [11:

OCH CN NCCH O To a suspension of 22.8 g of 2,6-dihydroxy-9-oxoxanthene in 200 ml of ethanol was added a solution of 4.6 g of sodium in 300 ml of ethanol. The solution was evaporated to dryness in a rotary evaporator and the residue was dissolved in 200 ml of N,N-dimethylforrnamide. After the addition of 23.8 g bromoacetonitrile the mixture was heated under reflux for 1 hour and then diluted with 300 ml of water. The precipitate thus formed was collected and a small amount was recrystallized from N,N-dimethylacetamide affording the analytical sample having a melting point of 218-223.

Anal. Calcd. for C I-1 N C, 66.66; H, 3.29; N, 9.15.

Found: C, 66.50; H, 3.35; N, 8.50.

EXAMPLE I] The following illustrates the preparation of 2-2 [(9- oxoxanthene-2,6-diyl)dioxy]diacetamidoxime, a compound of fonnula IV:

heated on a steam bath for 2 hours and filtered. The filtrate was diluted with 300 ml of water and the resulting precipitate was collected and recrystallized from N,N- dimethylacetamide with the addition of ethanol to induce precipitation affording 3.8 g of product which decomposed at 245.

Based on the assumed molecular formula C I-1 N 0 it was calculated that the elemental analysis by weight would be 54.84 percent carbon, 4.33 percent hydrogen and 15.05 percent nitrogen. The product was analyzed and found to contain 54.70 percent carbon, 4.31 percent hydrogen and 15.06 percent nitrogen which confirmed the accuracy of the assumed formula. This may be expressed:

Anal. Calcd. for C,-,H N O C, 54.84; H, 4.33; N,

Found: C, 54.70; H, 4.31; N, 15.06.

When evaluated in the foregoing test for trichomonacidal activity the compound produced a 16 percent kill at 100 micrograms per milliliter and a 33 percent kill at l milligram per milliliter.

What is claimed is:

l. A compound selected from those having the formula OCH C :NOH) NH H -N (HON: )CCH To a mixture of 30.6 g of [(9-oxoxanthene-2,6-diyl)- dioxyldiacetonitrile and 27.6 g of hydroxylamine hydrochloride in 300 ml of 1}],N-dimethylfo'rmarhide was added 53.0 g of sodium carbonate. The mixture was 

1. A COMPOUND SELECTED FROM THOSE HAVING THE FORMULA
 2. A compound as defined in claim 1 which is 2-2''-((9-oxoxanthene-2,6-diyl)dioxy)diacetamidoxime. 